Methods for treatment of beta-amyloid protein-induced ocular disease

ABSTRACT

The invention provides methods for treating beta-amyloid protein-involved ocular disease including age-related macular degeneration and glaucoma, pharmaceutical compositions and compounds useful for the same, and the use of these compounds for the manufacture of a medicament for treating the same. More particularly, the invention relates to the use of natural product compounds isolated from  Curcuma  sp.,  Zingiber  sp.,  Ginkgo biloba, Salvia  sp., and  Rosmarinus  sp. and synthetic chemical analogues thereof, for the treatment of a beta-amyloid protein-involved ocular disease.

RELATED APPLICATIONS

This application is a Continuation-in-Part of U.S. patent applicationSer. No. 11/128,638 filed May 13, 2005 which is a Continuation of U.S.patent application Ser. No. 11/084,316 filed Mar. 18, 2005 which is aContinuation of U.S. patent application Ser. No. 10/111,039 filed Apr.19, 2002 now U.S. Pat. No. 6,887,898 which was the U.S. National Phaseof International Patent Application Serial No. PCT/US00/41436 filed Oct.23, 2000, which claims priority from European Patent Application No.00986827.4, filed Oct. 23, 2000, which claims benefit of U.S.Provisional Patent Application Ser. No. 60/161,145 filed Oct. 22, 1999the disclosures of which are hereby incorporated by reference. Thisapplication also claims benefit of U.S. Provisional Patent ApplicationSer. No. 60/690,812 filed Jun. 15, 2005 the disclosure of which ishereby incorporated by reference. This application also claims benefitof U.S. Provisional Patent Application Serial No. ______ filed Nov. 23,2005 [Attorney Docket No. 30443/41465] entitled “SynergisticPharmaceutical Compositions Useful in Prevention and Treatment ofBeta-Amyloid Protein-Induced Disease Including Sage and Rosemary DerivedCompounds” the disclosure of which is hereby incorporated by reference.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The invention relates to the use of natural product compounds isolatedfrom plants, and synthetic chemical analogues thereof, for theprevention and treatment of beta-Amyloid protein-induced disease.Specifically, the invention relates to pharmaceutical compositions thatprotect neuronal cells from beta-Amyloid insult for use in preventingand treating beta-Amyloid protein-induced disease. More particularly theinvention is directed to protection of retinal cells and other visualfunction related cells of the eyes from beta-amyloid involved pathology,in particular, glaucoma and age-related macular degeneration (AMD).

2. Description of Related Technology

Alzheimer's disease (AD) is the most common cause of progressivecognitive dysfunction. AD affects approximately four million Americansand causes more than 100,000 deaths each year, with a total annual costapproaching $100 billion. It is estimated that by the year 2020, 14million Americans will be afflicted by the disease. See Carr et al., AmJ Med 103, 3S (1997) and Shastry, Am J Med Sci 315, 266 (1998).Furthermore, AD has a profound effect on the millions of family membersand other loved ones who provide most of the care for people having thisdisease. Unfortunately, the cure for AD has not yet been discovered.

The principal pathological characteristics of AD are senile plaques andneurofibrillary tangles (NTFs). Senile plaques are extracellulardeposits principally composed of insoluble aggregates of beta-amyloid(βA), that are infiltrated by reactive microglia and astrocytes. SeeSeidl et al., Neurosci. Lett 232, 49 (1997), Yan et al., Nature 382, 685(1997), Goedert, Trends Neurosci 16, 460 (1993), Haass et al., Cell 7,1039 (1994), Trojanowski et al., Am J Pathol 144, 449 (1994), Davis etal., Biochem Biophys Res Commun 189, 1096 (1992), Pike et al.,Neuroscience 13, 1676 (1993), Hensley et al., Proc Natl Acad Sci USA 91,3270 (1994), Behl et al., Cell 77, 817 (1994), Meda et al., Nature 374,647 (1995), and Klegeris et al., Biochem Biophys Res Commun 199, 984(1994). Plaques are diffusely distributed throughout the cerebral cortexof AD patients, and are the neuropathologic hallmark of the disease. SeeSeidl et al., Neurosci Lett 232, 49 (1997), Yan et al., Nature 382, 685(1997), Goedert, Trends Neurosci 16, 460 (1993), Haass et al., Cell 7,1039 (1994) and Trojanowski et al., Am J Pathol 144, 449 (1994). Theseplaques or PA fibril deposits are believed to be responsible for thepathology of a number of neurodegenerative diseases including, but notlimited to, Alzheimer's disease. NTFs are intraneuronal accumulation ofpaired helical filaments composed mainly of an abnormal form of tauprotein, that is a microtubule associated phosphoprotein which canpromote microtubule formation. See Goedert, Trends Neurosci 16, 460(1993), Haass et al., Cell 7, 1039 (1994) and Trojanowski et al., Am JPathol 144, 449 (1994). In the AD brain, the tau protein in NFTs ishyperphosphorylated (See Ihara et al., J Biochem 99, 1807 (1986)), acondition which has been suggested to contribute to the destabilizationof microtubule network, thereby impairing axonal network, and eventuallycausing neuronal death. See Trojanowski et al., FASEB J 9, 1570 (1995).NTFs occur primarily in medial temporal lobe structures (hippocampus,entorhinal cortex, and amygdala), and NTFs density appears to correlatewith dementia severity.

Senile plaques and NTFs appear to be involved in cerebral amyloidangiopathy, consequent neuronal loss, and cerebral atrophy leading todementia. Although research findings suggest that both plaques and NTFsare involved in disrupting nerve cell functions, the mechanisms thatlead to the pathology are not clearly understood.

βA has been suggested as one of the major causes of AD. βA was shown toexert direct toxic effects on neurons and to inhibit neurite growth invitro in a dose dependent manner. Thus, therapeutic approaches that canmodulate βA toxicity have been hypothesized to represent importantmethods for controlling the onset of AD. It is envisioned that ifneuronal cells can be protected from βA/senile plaque-induced toxicity,the onset of AD may be delayed or prevented. Current pharmacologicalapproaches related to AD preventive and neuroprotective interventionsinclude antioxidant therapy (See Lucca et al., Brain Res 764, 293(1997), Pike et al., J Neurochem 69, 1601 (1997), Manelli et al., BrainRes Bull 38, 569 (1995), Parnetti et al., Drugs 53, 752 (1997), Zhou etal., J Neurochem 67, 1419 (1996), Kumar et al., Int J Neurosci 79, 185(1994), Preston et al., Neurosci Lett 242, 105 (1998), and Tatton etal., Neurology 47, S171 (1996)), acetylcholinesterase inhibitors (SeeHoshi et al., J Biol Chem 272, 2038 (1997), Maurice et al., Brain Res706, 181 (1996), Harkany et al., Brain Res 695, 71 (1995), and Lahiri etal., J Neurosci Res 37, 777 (1994)), nicotinic and muscarinic agonists(See Maurice et al., Brain Res 706, 181 (1996), and Kihara et al., BrainRes 792, 331 (1998)), estrogen (See Ihara et al., J Biochem 99, 1807(1986), Henderson, Neurology 48 (5 Suppl. 7), S27 (1997), and Green etal., Neuroscience 84, 7 (1998)), nerve growth factor (NGF) (See Hefti,Neurobiol Aging 15 (Suppl 2), S193 (1994), and Seiger et al., BehavBrain Res 57, 255 (1993)), calcium channel blockers (See Zhou et al., JNeurochem 67, 1419 (1996) and Friedlich et al., Neurobiol Aging 15, 443(1994)), Zinc (See Cuajungco et al., Neurobiol Dis 4, 137 (1997)),sulfonated compounds (See Pollack et al., Neurosci Lett 197 211 (1995)and Lorenzo, et al., Ann NY Acad Sci 777, 89 (1996)), triaminopyridinenonopiate analgesic drug (See Muller et al., J Neurochem 68, 2371(1997)), low molecular lipophilic compounds that can activateneurotrophic factor signaling pathway (See Mattson, Neurosci BiobehavRev 21, 193 (1997)), and non-steroidal anti-inflammatory drugs such asibuprofen and aspirin (See Parnetti et al., Drugs 53, 752 (1997), Beardet al., Mayo Clin Proc 73, 951 (1998), and Pasinetti et al.,Neuroscience 87, 319 (1998)). Of particular interest to the presentinvention is the observation that an anti-βA protein antibody was shownto clear senile plaques and protect mutant PDAPP mice from the onset ofAD. See St George-Hyslop et al., Nature 400, 116 (1999).

The generation of reactive oxygen intermediates (ROS) through oxidativestress caused by βA has been suggested to be the major pathway of theβA-induced cytotoxicity. See Klegeris et al., Biochem Biophys Res Comun199, 984 (1994) and Lucca et al., Brain Res 764, 293 (1997). Senileplaques have been shown to exert a cytotoxic effect on neurons bystimulating microglia to produce reactive oxygen species (ROS). SeeSeidl et al., Neurosci Lett 232, 49 (1997), Yan et al., Nature 382, 685(1997), Goedert, Trends Neurosci 16, 460 (1993), Haass et al., Cell 7,1039 (1994), Trojanowski et al., Am J Pathol 114, 449 (1994), Davis etal., Biochem Biophys Res Commun 189, 1096 (1992), Pike et al.,Neuroscience 13, 1676 (1993), Hensley et al., Proc Natl Acad Sci USA 91,3270 (1994), Behl et al., Cell 77, 817 (1994), Meda et al., Nature 374,647 (1995) and Klegeris et al., Biochem Biophys Res Commun 199, 984(1994). The damaging effect of ROS can be prevented by the free radicalscavenging enzyme superoxide dismutase (SOD). See Thomas et al., Nature380, 168 (1996) and Manelli et al., Brain Res Bull 38, 569 (1995).

Aging of synthetic βA for 7 to 14 days at 37° C. in modified Eagle'smedia was also demonstrated to cause neurotoxic free radical formation.See Friedlich et al., Neurobiol Aging 15, 443 (1994) and Puttfarcken etal., Exp Neurol 138, 73 (1996). However, aging PA in the presence of themedia supplement B27, which contains antioxidants as well as otheragents that provide protection against oxidative damage, has been shownto inhibited βA-induced neurotoxicity. See Thomas et al., Nature 380,168 (1996) and Manelli et al., Brain Res Bull 38, 569 (1995).

In addition to βA peptide-induced ROS mediated neurotoxicity, βA peptidehas been shown to cause neuronal cell death by stimulating microglialexpression of tumor necrosis factor β (TNFβ). See Tarkowski et al.,Neurology 54, 2077 (2000) and Barger et al., Proc Natl Acad Sci USA 92,9328 (1995). The accumulation of βA peptide as neuritic plaques is knownto be both trophic and toxic to hippocampal neurons, causing apoptosisor necrosis of the neurons in a dose dependent manner. βA peptide wasdemonstrated to induce these cellular effects by binding with a receptorfor advanced glycation end products (RAGE) that was previously known asa central cellular receptor for advanced glycation endproducts. SeeArancio et al., EMBO J23, 4096 (2004), Huttunen et al., J Biol Chem 274,19919 (1999), and Yan et al., Nature 382, 685 (1996). RAGE was suggestedto mediate the interaction of PA peptide with neurons and withmicroglia, resulting in oxidative stress mediated cytotoxicity. BlockingRAGE with anti-RAGE F(αβ′)₂ prevented the appearance of TNFβ messengerRNA and diminished TNFβ antigen to levels seen in untreated cells. Thus,it is postulated that RAGE mediates microglial activation by βA peptideby producing cytotoxic cytokines that cause neuronal damage in ADpatients. In addition, RAGE was also demonstrated to specifically bindwith βA peptide and mediate βA peptide-induced oxidative stress.

Cell receptors that bind to βA peptide have been identified. Thelow-affinity neurotrophin receptor p75 (p75NTR) which belongs to thefamily of apoptotic receptors that generate cell-death signals onactivation was found throughout the brains of AD patients. βA peptidewas found to be a ligand for p75NTR, and to cause preferential apoptosisof neurons and normal neural crest-derived melanocytes that expressp75NTR upon specifically binding to p75NTR. See Zhang et al., J Neurosci23, 7385 (2003) and Perini et al., J Exp Med 195, 907 (2002).

Basal forebrain cholinergic neurons express the highest levels of p75NTRin the adult human brain and have been shown to be involved in AD. Theexpression of p75NTR neuronal cells was shown to potentiate βApeptide-induced cell death. This interaction of βA peptide with p75NTRto mediate neuronal death in AD suggested a new target for therapeuticintervention. See Zhang et al., J Neurosci 23, 7385 (2003) and Perini etal., J Exp Med 195, 907 (2002).

Recently, ERAB which is over-expressed in neurons of the AD brain, wasshown to bind with βA peptide to induce neuronal death in AD. BlockingERAB with an antibody, anti-ERAB F(ab′)₂, was found to reduce the βApeptide-induced cell death while ERAB overexpression increases βApeptide-induced cell death. See Frackowiak et al., Brain Res 907, 44(2001) and Yan et al., J Biol Chem 274, 2145 (1999).

In designing inhibitors of βA peptide toxicity, it was found thatneither the alteration of the apparent secondary structure of βA peptidenor the prevention of βA peptide aggregation is required to abrogate thecytotoxicity of βA peptide. Nonetheless, inducing changes in aggregationkinetics and in higher order structural characteristics of βA peptideaggregates also proved to be effective in reducing βA peptide toxicity.See Soto et al., Neuroreport 7, 721 (1996). Synthetic inhibitors thatinteract with βA peptide were shown to completely block βA peptidetoxicity against PC12 cells, demonstrating that complete disruption ofamyloid fibril formation is not necessary for abrogation of toxicity. Itwas also demonstrated that dipolar compounds such as phloretin andexifone that decrease the effective negative charge of membranes canprevent the association of βA peptide with negatively charged lipidvesicles and thereby prevent βA peptide-induced cytotoxicity. See Hertelet al., Proc. Natl. Acad. Sci. USA 94, 9412 (1997). These resultssuggest that PA peptide toxicity can be mediated through aphysicochemical interaction with cell membranes.

Glaucoma and age-related macular degeneration (AMD) are the most commonleading cause of irreversible progressive visual dysfunction that leadsto blindness. Glaucoma causes irreversible vision loss worldwide anestimated 66.8 million people. See Khaw et al., BMJ 320, 1619 (2000).AMD is the leading cause of blindness and vision loss in developingcountries due to increased life expectancy and subsequent increase inaged population. See VanNewkirk et al., Ophthalmol 108, 960 (2001).Between 20 and 25 million people are affected by AMD worldwide, a figurethat will triple with the increase in the aging population in the next30˜40 years. See Smith et al., Ophthalmol 108, 697 (2001) and McCarty etal., Arch Ophthalmol 119, 1455 (2001). There are over 200,000 cases ofneovascular degeneration that present to ophthalmologist in the UnitedStates each year. See Bressler et al., BMJ 321, 1425 (2000) and Chopdaret al., BMJ 326, 485 (2003). Glaucoma and AMD have profound effect onthe family members and other loved ones who provide most of the care forpeople having this disease. Unfortunately, the cure for glaucoma and AMDhas not yet been discovered.

AMD is characterized by abnormal extracellular deposits, known asdrusen, the hallmark sign of AMD, that accumulate along the basalsurface of the retinal pigmented epithelium. Although drusen is commonin older individuals, large numbers of drusen and/or extensive areas ofconfluent drusen represent a significant risk factor for AMD. Widespreaddrusen deposition is associated with retinal pigmented epithelial celldysfunction and degeneration of the photoreceptor cells. See Johnson etal., Proc Natl Acad Sci USA 99, 11830 (2002). There are two types ofAMD, dry and wet. The dry type of AMD is characterized by a geographicatrophy that progresses slowly over many years. In the wet type of AMDchoroidal neovascularization occurs that result in a dense fibrovascularscar that may involve the entire macular area. The wet type of AMD ismore sight threatening than the dry type and is responsible for 90% ofcases of severe visual loss in elderly population. See Chopdar et al.,BMJ 326, 485 (2003).

Glaucoma is a chronic neurodegeneration of the optic nerve, retinalganglion cells, that result in irreversible vision loss. See Khaw etal., BMJ 320, 1619 (2000).

The pathogenesis of glaucoma and AMD has recently been linked todeposition of beta-amyloid (βA) in retinal cells of the eyes. It wasrecently demonstrated that retinal ganglion cell death in glaucomainvolves βA neurotoxicity at the molecular level. See McKinnon et al.,IOVS 43, 1077 (2002). βA was also shown to associate with asubstructural vesicular component within drusen and was found tocorrelate with the location of degenerating photoreceptors and retinalpigmented epithelium cells. See Dentchev et al., Mol Vis 14, 184 (2003).βA deposition was found an important component of the local inflammatoryevents that contribute to atrophy of the retinal pigmented epithelium,drusen biogenesis and the pathogenesis of AMD. See Johnson et al., ProcNatl Acad Sci USA 99, 11830 (2002).

Thus, therapeutic approaches that can modulate βA toxicity have beenhypothesized to represent important methods for controlling the onset ofglaucoma and macular degeneration. It is envisioned that if retinalcells can be protected from βA-induced toxicity, the onset of glaucomaand AMD may be delayed or prevented.

Current glaucoma treatment focuses on lowering intraocular pressure, themajor risk factor for the disease. Glaucoma has been treated medically,surgically, or with laser to lower intraocular pressure that can slowthe disease progression. Pharmacological treatment approaches are:cholinergic agents (pilocarpine—increases outflow of the aqueous humour;beta blockers—reduce aqueous secretion), oral carbonic anhydraseinhibitors (acetazolamide and dorzolamide—reduces aqueous secretion),alpha-2 adrenergic agonists (apraclonidine and brimonidine), andprostaglandin agonists (latanoprost—opens up an alternative pathway foraqueous outflow by altering the resistance of the extracellular matrix).See Khaw et al., BMJ 320, 1619 (2000) and Khaw et al., BMJ 328, 156(2004).

One current treatment approach for AMD is a technique calledphotodynamic therapy that uses verteporfin as the photosensitizer. Longterm supplementation with high dose zinc and antioxidant vitamins (A, C,and E) showed a significant reduction in the relative risk of developingneovascular AMD. As a preventive measure against the disease progressionand the onset of AMD, carotenoids lutein and zeaxanthin, which arepotent antioxidants found in high concentrations in the macular retinaare found to be effective. See. Chopdar et al., BMJ 326, 485 (2003).

One important pharmacological approach related to PA-inducedneurodegenerative disease preventive and neuroprotective interventionsmay be antioxidant therapy. See Kumar et al., Int J Neurosci 79, 185(1994), Lucca, et al., Brain Res 764, 293 (1997), Manelli et al., BrainRes Bull 38, 569 (1995), Parnetti et al., Drugs 53, 752 (1997), Prestonet al., Neurosci Lett 242, 105 (1998), and Zhou, et al., J Neurochem 67,1419 (1996). In designing inhibitors of βA toxicity, it was found thatinducing changes in aggregation kinetics and in higher order structuralcharacteristics of βA aggregate may prove to be effective in reducing βAtoxicity. See Ghanta et al., J Biol Chem 271, 29525 (1996). Syntheticinhibitors that interact with βA was shown to completely block βAtoxicity against PC12 cells, demonstrating that complete disruption ofamyloid fibril formation is not necessary for abrogation of toxicity.See Yaar et al., J Clin Invest 100, 2333 (1997) and Hertel et al., ProcNatl Acad Sci USA 94, 9412 (1997). These results suggest that βAtoxicity can be mediated through a physicochemical interaction with cellmembranes.

There is strong interest in discovering potentially valuable naturalsources for drug development. One reasonable source of such naturalproducts involves medicinal plants that have been in use throughouthistory for treating various ailments. Thus, the discovery ofpotentially valuable plants that can protect neurons from βA insult isof interest.

Curcuma longa (Zingiberaceae) has been used as curry spice and a wellknown constituent of Indonesian traditional medicine. See Nurfina etal., Eur J Med Chem 32, 321 (1997). One of the important constituents ofturmeric is curcumin that has been known as a natural antioxidant withantitumor activity. See Ruby et al., Cancer Lett 94, 79 (1995). Fromturmeric, curcuminoids with antioxidant property have been demonstratedto protect neuronal cells from βA insult. See Kim DSHL et al., NeurosciLett 303, 57 and Park S Y et al., J Nat Prod 65, 1227 (2002). Arepresentative list of Curcuma sp. include C. longa, C. aromatica, C.domestica, C. xanthorrhiza, and C. zedoaria.

Zingiber officinale (Zingiberaceae) is one of the world's favoritespices, probably discovered in the tropics of Southeast Asia. Ginger hasbenefited humankind as a wonder drug since the beginning of recordedhistory. See Jitoe et al., J Agric Food Chem 40, 1337 (1992), Kikuzakiet al., J Food Sci 58, 1407 (1993) and Schulick, Herbal Free Press, Ltd.(1994). From ginger, shogaols with antioxidant property have also beendemonstrated to protect neuronal cells from βA insult. See Kim et al.,Planta Medica 68, 375 (2002). A representative list of Zingiber sp.include Z. officinale, Z. zerumbet, and Z. mioga.

Ginkgo (Ginkgo biloba (Ginkgoaceae)) is an herbal that has been used totreat neurologic ailment for thousand years as an Asian traditionalmedicine. Ginkgo leaf extract has shown to exhibit potent antioxidantactivity and are widely used in the dietary supplement industry. Theantioxidant activity of ginkgo has shown to be primarily contributed byditerpenes such as ginkgolides, bilobilide, flavonoids, and ginkgolicacids. See Hopia et al., J Agric Food Chem 44, 2030 (1996) and Nakataniet al., Agric Biol Chem 47, 353 (1983).

Sage (Salvia officinalis L. (Lamiaceae)) and Rosemary (Rosmarinusofficinalis L. (Labiatae)) are spices widely used for flavoring andseasoning foods. These spices have shown to contain potent diterpenoidantioxidants such as carnosic acid, carnosol, rosmarinic acid, rosmanol,epirosmanol, rosmadial, isorosmanol etc. See Haraguchi et al., PlantaMed 61, 333 (1995). Inatani et al, Agric Biol Chem 47: 521 (1983).Nakatani et al., Agric Biol Chem 48: 2081 (1984). Inatani et al., AgricBiol Chem 46: 1661 (1982). Wang et al., J Agric Food Chem 46: 2509(1998). Wang et al., JAgric Food Chem 46: 4869 (1998).

SUMMARY OF THE INVENTION

The present invention relates to the identification and isolation ofnatural compounds present in turmeric, ginger, gingko biloba, sage, androsemary that exhibit potent anti-βA peptide activity. The inventionfurther provides novel synthetic compounds exhibiting potent anti-βApeptide activity which includes but is not limited to, the ability toneutralize amyloid protein mediated cytotoxicity towards retinal cellsthat relate to the pathogenesis of glaucoma and AMD. Specifically, theinvention provides compounds and pharmaceutical compositions capable ofprotecting neurons from βA peptide insult, and methods for treating βAprotein-induced disease with the same. In addition, it has been foundthat compounds derived from sage and rosemary and their analogs andhomologs as described in co-owned and copending U.S. Provisional PatentApplication Ser. No. ______ filed Nov. 23, 2005 [Attorney Docket No.30443/41465] entitled “Synergistic Pharmaceutical Compositions Useful inPrevention and Treatment of Beta-Amyloid Protein-Induced DiseaseIncluding Sage and Rosemary Derived Compounds” the disclosure of whichis hereby incorporated by reference. These compounds have potentanti-Beta-amyloid activity alone and may be combined with the otherturmeric, ginger, and ginkgo-biloba derived compounds and their analogsand homologues have anti Beta-amyloid activity to treat beta-Amyloidprotein-induced ocular disease including age-related maculardegeneration (AMD) and glaucoma.

As used herein, synthetic turmeric, ginger, ginkgo biloba, sage, orrosemary compounds include chemically synthesized versions of naturallyoccurring turmeric sp., ginger sp., ginko biloba, sage sp., or rosemarysp. compounds respectively as well as analogues and homologues of suchnaturally occurring compounds which have anti-βA peptide activity. Asused herein anti-βA peptide activity includes, but is not limited to,the ability to neutralize amyloid protein mediated cytotoxicityincluding neurotoxicity.

Thus, the present invention is directed to treating (which when usedherein also includes preventing) βA-involved ocular disease glaucoma andAMD. According to one aspect of the invention, an extract containingnatural compounds found in turmeric (as well as synthetic analogues andhomologues thereof) as the major ingredients or components and thenatural compounds found in turmeric (as well as synthetic analogues andhomologues thereof), may be administered to protect retinal cells fromβA-involved cytotoxicity. Natural compounds that are suitable for usewith the invention include, but are not limited to4″-(3′″-methoxy-4′″-hydroxyphenyl)-2″-oxo-3″-enebutanyl3-(3′-methoxy-4′-hydroxyphenyl)propenoate (calebin-A) and1,7-bis(4-hydroxy-3-methoxyphenyl)-1,4,6-heptatrien-3-one, and sevenknown compounds,1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione (curcumin),1-(4-hydroxy-3-methoxyphenyl)-7-(4-hydroxyphenyl)-1,6-heptadiene-3,5-dione(demethoxycurcumin), 1,7-bis(4-hydroxyphenyl)-1,6-heptadiene-3,5-dione(bisdemethoxycurcumin),1-hydroxy-1,7-bis(4-hydroxy-3-methoxyphenyl)-6-heptene-3,5-dione,1,7-bis(4-hydroxyphenyl)-1-heptene-3,5-dione,1,7-bis(4-hydroxyphenyl)-1,4,6-heptatrien-3-one, and1,5-bis(4-hydroxy-3-methoxyphenyl)-1,4-pentadien-3-one, 2-shogaol,4-shogaol, 6-shogaol, 8-shogaol, 10-shogaol, 12-shogaol, 2-gingerol,4-gingerol, 6-gingerol, 8-gingerol, 10-gingerol, 12-gingerol, ginkgolicacids, rosmanol, isorosmanol, rosmadial, carnosol, carnosic acid,epirosmanol, rosmarinic acid etc.

In one aspect, the invention relates to a method for the treatment of abeta-Amyloid protein-induced disease including but not limited toAlzheimer's Disease (AD), age-related macular degeneration (AMD) andglaucoma comprising administering to a subject suffering from thebeta-Amyloid protein-induced disease a therapeutically effective amountof a compound having the formula (I):

In this formula, the dotted configuration is optionally a single bond ora double bond. Generally, R₁ is selected from the group consisting ofOH, OMe, OR₅₀, and X wherein R₅₀ is alkyl, alkenyl, or alkynyl, and X isF, Cl, Br, or I. Preferably, R₁ is selected from the group consisting ofOH, OMe, OR₅₀ and X wherein R₅₀ is (CH₂)_(n)CH₃ and n is 1-7 and X is F,Cl, Br, or I. More preferably, R₁ is selected from the group consistingof H, OH, and OMe. Even more preferably, R₁ is OH. Even more preferably,R₁ is selected from the group consisting of H and OMe when the dottedconfiguration of compound (I) is a double bond, and R₁ is selected fromthe group consisting of H and OH when the dotted configuration is asingle bond. Generally, R₂ is selected from the group consisting of OH,OMe, OR₅₀, and X wherein R₅₀ is alkyl, alkenyl, or alkynyl, and X is F,Cl, Br, or I. Preferably, R₂ is selected from the group consisting ofOH, OMe, OR₅₀ and X wherein R₅₀ is (CH₂)_(n)CH₃ and n is 1-7 and X is F,Cl, Br, or I. More preferably, R₂ is selected from the group consistingof H, OH, and OMe. Even more preferably, R₂ is OH. Even more preferably,R₂ is selected from the group consisting of H and OMe when the dottedconfiguration of compound (I) is a double bond, and R₂ is H when thedotted configuration is a single bond.

Other compounds useful for practice of the invention include those ofthe formula (II):

In this formula, the dotted configuration is optionally a single bond ora double bond or a triple bond. Generally, R₃ is selected from the groupconsisting of OH, OMe, OR₅₀, and X wherein R₅₀ is alkyl, alkenyl, oralkynyl, and X is F, Cl, Br, or I. Preferably, R₃ is selected from thegroup consisting of OH, OMe, OR₅₀ and X wherein R₅₀ is (CH₂)_(n)CH₃ andn is 1-7 and X is F, Cl, Br, or I. More preferably, R₃ is selected fromthe group consisting of H, OH, and OMe. Even more preferably, R₃ is H.Generally, R₄ is selected from the group consisting of OH, OMe, OR₅₀,and X wherein R₅₀ is alkyl, alkenyl, or alkynyl, and X is F, Cl, Br, orI. Preferably, R₄ is selected from the group consisting of OH, OMe, OR₅₀and X wherein R₅₀ is (CH₂)_(n)CH₃ and n is 1-7 and X is F, Cl, Br, or I.More preferably, R₄ is selected from the group consisting of H, OH, andOMe. Even more preferably, R₄ is H. Even more preferably, R₄ is H whenthe first dotted configuration of compound (II) is a double bond and thesecond dotted configuration of compound (II) is a single bond, R₄ is Hwhen both dotted configurations are single bonds, and R₄ is selectedfrom the group consisting of H and OMe when both dotted configurationsare double bonds. Generally, R₅ is selected from the group consisting ofH, OH, OMe, OR₅₀, and X wherein R₅₀ is alkyl, alkenyl, or alkynyl, and Xis F, Cl, Br, or I. Preferably, R₅ is selected from the group consistingof H, OH, OMe, OR₅₀, and X wherein R₅₀ is (CH₂)_(n)CH₃ and n is 1-7, ansX is F, Cl, Br, or I. More preferably, R₅ is selected from the groupconsisting of H, OH, and OMe. Even more preferably, R₅ is OH.

While compounds of formula (II) have been presented herein as diketones,and compounds of formula (I) have been presented as enols, those ofskill in the art recognize that diketones and enols can coexist insolution as tautomers as shown below.

Accordingly, the invention contemplates the use and production ofcompounds in either tautomeric form, and as a mixture of the two forms.

A natural product compound having the following general formula wasisolated from turmeric, and was found to protect cells from βApeptide-induced toxicity.

Still other turmeric-related compounds useful in practice of theinvention include those of formula (III):

In this formula, the dotted configuration is optionally a single bond ora double bond or a triple bond. Z is a representation of isostericvariation in which Z is selected from O, S, NH, NR₆₀, where R₆₀ isalkyl, alkenyl, or alkynyl. Generally, R₆ is selected from the groupconsisting of OH, OMe, OR₅₀, and X wherein R₅₀ is alkyl, alkenyl, oralkynyl, and X is F, Cl, Br, or I. Preferably, R₆ is selected from thegroup consisting of OH, OMe, OR₅₀ and X wherein R₅₀ is (CH₂)_(n)CH₃ andn is 1-7 and X is F, Cl, Br, or I. More preferably, R₆ is selected fromthe group consisting of OH and OMe. Even more preferably, R₆ is OH.Generally, R₇ is selected from the group consisting of OH, OMe, OR₅₀,and X wherein R₅₀ is alkyl, alkenyl, or alkynyl, and X is F, Cl, Br, orI. Preferably, R₇ is selected from the group consisting of OH, OMe, OR₅₀and X wherein R₅₀ is (CH₂)_(n)CH₃ and n is 1-7 and X is F, Cl, Br, or I.More preferably, R₇ is selected from the group consisting of H, OH, andOMe. Even more preferably, R₇ is H. Generally, R₈ is selected from thegroup consisting of OH, OMe, OR₅₀, and X wherein R₅₀ is alkyl, alkenyl,or alkynyl, and X is F, Cl, Br, or I. Preferably, R₈ is selected fromthe group consisting of OH, OMe, OR₅₀ and X wherein R₅₀ is (CH₂)_(n)CH₃and n is 1-7 and X is F, Cl, Br, or I. More preferably, R₈ is selectedfrom the group consisting of H, OH, and OMe. Even more preferably, R₈ isOH. Generally, R₉ is selected from the group consisting of OH, OMe,OR₅₀, and X wherein R₅₀ is alkyl, alkenyl, or alkynyl, and X is F, Cl,Br, or I. Preferably, R₉ is selected from the group consisting of OH,OMe, OR₅₀ and X wherein R₅₀ is (CH₂)_(n)CH₃ and n is 1-7 and X is F, Cl,Br, or I. More preferably, R₉ is selected from the group consisting ofH, OH, and OMe. Even more preferably, R₉ is H.

The second set of compounds useful for practice of the invention includenatural compounds which can be extracted on otherwise derived fromGinkgo biloba as well as synthetic Ginkgo biloba compounds includingbiologically active homologues and analogues of natural Ginkgo bilobacompounds which share anti-βA activity. Such compounds have the formula(IV):

or a pharmaceutically acceptable salt or ester thereof, wherein R isselected from the group consisting of higher alkyl, higher alkenyl, andhigher alkynyl.

More preferably, R is

and n is 1-7. Even more preferably, R is selected from the groupconsisting of

And R is also selected from the group consisting of alkyl, alkenyl, andalkynyl; for example;

and y is 1-9, or having more than one double bond (cis or trans), ortriple bond consisting of; for example;

wherein the dotted line configuration is optionally a single bond (cisor trans), or a triple bond, wherein the alkyl, alkenyl, and alkynylgroup is selected from ethers and/or thioethers or amines; for example;

wherein z=O, S, NR_(n), where R=alkyl, alkenyl, alynyl groups; and n=1or 2.

The third set of compounds useful for practice of the invention includenatural compounds which can be extracted on otherwise derived fromZingiber sp. (ginger) as well as synthetic ginger compounds includingbiologically active homologues and analogues of natural ginger compoundswhich share anti-βA activity. Such compounds have the formula (V):

In this formula, the dotted configuration is optionally a single bond ora double bond or a triple bond. Preferably, R₁₀ is selected from thegroup consisting of OH, OMe, OR′, and X wherein R′ is alkyl, alkenyl, oralkynyl, and X is F, Cl, Br, or I. More preferably, R₁₀ is selected fromthe group consisting of OH, OMe, OR″, and X wherein R″ is (CH₂)_(n)CH₃and n is 1-7, and X is F, Cl, Br, or I. Even more preferably, R₁₀ is OH.Preferably, R₁₁ is selected from the group consisting of H, OH, OMe, andOR′ wher R′ is alkyl, alkenyl, or alkynyl. More preferably, R₁₁ isselected from the group consisting of H, OH, OMe, and OR″ wherein R″ is(CH₂)_(n)CH₃ and n is 1-7. Even more preferably, R₁₁ is selected fromthe group consisting of H and OMe. Preferably, R₁₂ is selected from thegroup consisting of alkyl, alkenyl, and alkynyl. More preferably, R₁₂ isselected from the group consisting of

and y is 1-9. Even more preferably, R₁₂ is selected from the groupconsisting of

and y is 1-9, or having more than one double bond (cis or trans), ortriple bond consisting of; for example;

wherein the dotted line configuration is optionally a single bond (cisor trans), or a triple bond, wherein the alkyl, alkenyl, and alkynylgroup is selected from ethers and/or thioethers or amines; for example;

wherein z=O, S, NR_(n), where R=alkyl, alkenyl, alynyl groups; and n=1or 2.

And compounds having a formula (VI):

In this formula, the dotted configuration is optionally a single bond ora double bond or a triple bond. Preferably, R₁₃ is selected from thegroup consisting of OH, OMe, OR′, and X wherein R′ is alkyl, alkenyl, oralkynyl, and X is F, Cl, Br, or I. More preferably, R₁₃ is selected fromthe group consisting of OH, OMe, OR″, and X wherein R″ is (CH₂)_(n)CH₃and n is 1-7, and X is F, Cl, Br, or I. Even more preferably, R₁₃ is OH.Preferably, R₁₄ is selected from the group consisting of H, OH, OMe, andOR′ wher R′ is alkyl, alkenyl, or alkynyl. More preferably, R₁₄ isselected from the group consisting of H, OH, OMe, and OR″ wherein R″ is(CH₂)_(n)CH₃ and n is 1-7. Even more preferably, R₁₄ is selected fromthe group consisting of H and OMe. Preferably, R₁₅ is selected from thegroup consisting of alkyl, alkenyl, and alkynyl. More preferably, R₁₅ isselected from the group consisting of

and y is 1-9. Even more preferably, R₁₅ is selected from the groupconsisting of

and y is 1-9, or having more than one double bond (cis or trans), ortriple bond consisting of; for example;

wherein the dotted line configuration is optionally a single bond (cisor trans), or a triple bond, wherein the alkyl, alkenyl, and alkynylgroup is selected from ethers and/or thioethers or amines; for example;

wherein z=O, S, NR_(n), where R=alkyl, alkenyl, alynyl groups; and n=1or 2.

It is apparent from the biological results for the ginger-derivednatural product compounds that the length of the side chain is importantfor the expression of biological activity. For example, with respect tothe ginger-derived natural product compounds, compounds (11), (12), (13)and (14), the biological activity appears to improve as the compounds'side chain length increases. Thus, it is of interest to prepareanalogues having different and lengthier side-chains. Preferably,shogaol compounds have side chains wherein R₁₂ has five or more carbons.More preferably, R₁₂ has nine or more carbons, and even more preferably,R₁₂ has eleven or more carbons. Furthermore, two of the synthesizedshogaol analogue compounds, compounds (45) and (50), also effectivelyprotected cells from βA peptide insult despite the fact that thesecompounds have different substituents than the ginger-derived naturalproduct compounds. For example, compound (45) differs from theginger-derived natural product compounds because it has a saturatedhydrocarbon side chain, and compound (50) differs from theginger-derived natural product compounds because it does not have amethoxy substituent. These data suggest that changing the nature of thesubstituents on the phenyl rings of the active compounds is of interestfor the methods, pharmaceutical compositions, compounds and usesaccording to the invention.

As used herein, the term “alkyl” refers to a carbon chain having atleast two carbons. Preferably, alkyl refers to a carbon chain havingbetween two and twenty carbons. More preferably, alkyl refers to acarbon chain having between two and eight carbons. The term “alkenyl,”as used herein, refers to a carbon chain having at least two carbons,and at least one carbon-carbon double bond. Preferably, alkenyl refersto a carbon chain having between two and twenty carbons, and at leastone carbon-carbon double bond. More preferably, the term alkenyl refersto a carbon chain having between two and eight carbons, and at least onecarbon-carbon double bond. The term “alkynyl,” as used herein, refers toa carbon chain having at least two carbon atoms, and at least onecarbon-carbon triple bond. Preferably, alkynyl refers to a carbon chainhaving between two and twenty carbon atoms, and at least onecarbon-carbon triple bond. More preferably, alkynyl refers to a carbonchain having between two and eight carbon atoms, and at least onecarbon-carbon triple bond.

As used herein, the term “higher alkyl” refers to a carbon chain havingat least five carbon atoms. Preferably, higher alkyl refers to a carbonchain having between five and twenty carbons. More preferably, higheralkyl refers to a carbon chain having between five and twelve carbonatoms. As used herein, the term “higher alkenyl” refers to a carbonchain having at least five carbon atoms, and at least one cabon-carbondouble bond. Preferably, higher alkenyl refers to a carbon chain havingbetween five and twenty carbon atoms, and at least one carbon-carbondouble bond. More preferably, higher alkenyl refers to a carbon chainhaving between five and twelve carbon atoms, and at least onecarbon-carbon double bond. The term “higher alkynyl,” as used herein,refers to a carbon chain having at least five carbons, and at least onecarbon-carbon triple bond. Preferably, higher alkynyl refers to a carbonchain having between five and twenty carbon atoms, and at least onecarbon-carbon triple bond. More preferably, the term higher alkynylrefers to a carbon chain having between five and twelve carbon atoms,and at least one carbon-carbon triple bond.

The fourth set of compounds useful for practice of the invention includenatural compounds which can be extracted or otherwise derived fromSalvia sp. (sage) and Rosmarinus sp. (rosemary) which share anti-βAactivity. Such compounds have the formulas (VII), (VIII), and (IX):

According to one aspect of the invention, combinations of each of theanti-Beta amyloid compounds may be administered as well as combinationsof compounds of natural or synthetic compounds selected from thedifferent classes of tumeric, ginkgo biloba, ginger, sage and rosemarycompounds may be administered in combination for additive or synergisticeffect. The invention also provides methods whereby the plant-derivedcompounds and homologues and analogues thereof may be combined withother agents including those selected from the group consisting ofcholinergic agents (such as pilocarpine, beta blockers), oral carbonicanhydrase inhibitors (such as acetazolaminde and dorzolamide), alpha-2adrenergic agonists (such as apraclonidine and brimonidine),prostaglandin agonists (latanoprost), carotenoids, lutein andzeaxanthin.

BRIEF DESCRIPTION OF THE DRAWINGS

These and other features, aspects, and advantages of the presentinvention will become better understood with regard to the followingdescription, appended claims, and accompanying drawings where:

FIG. 1 shows the structures of turmeric-derived natural productcompounds that protected PC12, IMR32, and HUVEC cells from βApeptide-induced toxicity.

FIG. 2 shows a scheme for the synthesis of dihydro- andtetrahydro-curcuminoids.

FIG. 3 shows a scheme for the synthesis of symmetric and unsymmetriccurcumin analogues and related compounds.

FIG. 4 shows a scheme for the synthesis of turmeric-derived naturalproduct compound (6).

FIG. 5 shows the structures of curcuminoid compounds that have beensynthetically prepared and assayed for biological activity against βApeptide-induced toxicity.

FIG. 6 shows the structures of ginger-derived natural product compoundsthat protected PC12, IMR32, and HUVEC cells from βA peptide-inducedtoxicity.

FIG. 7 shows a scheme for the synthesis of ginger-derived naturalproduct compound (13).

FIG. 8 shows a scheme for the synthesis of [9]-dihydroshogaol, compound(45).

FIG. 9 shows a scheme for the synthesis of [9]-demothoxyshogaol,compound (50).

FIG. 10 shows the structures of ginkgo biloba-derived natural productcompounds that protected PC 12 and HUVEC cells from βA peptide-inducedtoxicity.

FIG. 11 shows a proposed synthesis for ginkolic acids and theiranalogues.

DETAILED DESCRIPTION OF THE INVENTION

One aspect of the present invention is directed to the use of methanoland other extract of Curcuma sp. (Zingiberaceae), Zingiber sp.(Zingiberaceae), Ginkgo biloba, Salvia sp. (Lamiaceae) and Rosmarinussp. (Labiatae) to effectively protect cells from PA insult. The extractis obtained by pharmacologically acceptable solvent that is comprised ofbut not limited to methanol, ethanol, isopropyl alcohol, butanol etc. ofsuch nature, and other nonalcoholic solvents such as dimethylsulfoxide,dimethyl formate, chloroform, dichloromethane, hexanes, petroleum etherand diethyl ether types, and in combination with water. The extracts ofthese plants were found to protect PC12, IMR32, and HUVEC cells from βAinsult. Via bio-assay guided fractionation, twelve natural productcompounds (eleven known and one novel) exhibiting potent anti-βA peptideactivity were isolated and identified. These natural product compoundswere found to protect PC12, IMR32, HUVEC, and primary cortical ratneuronal cells from βA peptide (both 25-35 and 1-42) insult.

In some cases, the natural product compounds were syntheticallyprepared. It is necessary and cost efficient to chemically synthesizethe compounds in order to perform a thorough bioassay because only asmall amount of these compounds are available from the natural sources.The biological activities of the synthesized natural product compoundswere identical to those of the natural product compounds isolated fromthe plants. A series of natural product analogues that protect cellsfrom βA peptide insult as effectively as the isolated natural productcompounds were also synthesized.

Methods of treating a beta-Amyloid protein-induced ocular diseaseincluding AD, AMD and glaucoma with the compounds of the invention aredescribed herein. Further, pharmaceutical compositions comprising one ormore compounds of the invention and a pharmaceutically acceptablediluent, adjuvant, or carrier are provided. The use of the compounds ofthe invention for the manufacture of a medicament for treatment of abeta-amyloid protein-induced ocular disease is also disclosed herein.

Natural product compounds having the following general formula wereisolated from turmeric and were found to protect cells from βA peptideinsult. In addition, several of the natural product compounds describedby this general formula were synthetically prepared.

In this formula, the dotted configuration is optionally a single bond ora double bond. Generally, R₁ is selected from the group consisting of H,OH, OMe, and OR₅₀ wherein R₅₀ is alkyl, alkenyl, or alkynyl. Preferably,R₁ is selected from the group consisting of H, OH, OMe, and OR₆₀ whereinR₆₀ is (CH₂)_(n)CH₃ and n is 1-7. More preferably, R₁ is selected fromthe group consisting of H, OH, and OMe. Even more preferably, R₁ isselected from the group consisting of H and OMe when the dottedconfiguration of compound (I) is a double bond, and R₁ is selected fromthe group consisting of H and OH when the dotted configuration is asingle bond. Generally, R₂ is selected from the group consisting of H,OMe, and OR₅₀ wherein R₅₀ is alkyl, alkenyl, or alkynyl. Preferably, R₂is selected from the group consisting of H, OMe, and OR₆₀ wherein R₆₀ is(CH₂)_(n)CH₃ and n is 1-7. More preferably, R₂ is selected from thegroup consisting of H and OMe. Even more preferably, R₂ is selected fromthe group consisting of H and OMe when the dotted configuration ofcompound (I) is a double bond, and R₂ is H when the dotted configurationis a single bond.

Other compounds useful for practice of the invention include those ofthe formula (II):

In this formula, the dotted configuration is optionally a single bond ora double bond or a triple bond. Generally, R₃ is selected from the groupconsisting of H, OMe, and OR₅₀ wherein R₅₀ is alkyl, alkenyl, oralkynyl. Preferably, R₃ is selected from the group consisting of H, OMe,and OR₆₀ wherein R₆₀ is (CH₂)_(n)CH₃ and n is 1-7. More preferably, R₃is selected from the group consisting of H and OMe. Even morepreferably, R₃ is H. Generally, R₄ is selected from the group consistingof H, OH, OMe, and OR₅₀ wherein R₅₀ is alkyl, alkenyl, or alkynyl.Preferably, R₄ is selected from the group consisting of H, OH, OMe, andOR₆₀ wherein R₆₀ is (CH₂)_(n)CH₃ and n is 1-7. More preferably, R₄ isselected from the group consisting of H, OH, and OMe. Even morepreferably, R₄ is H when the first dotted configuration of compound (II)is a double bond and the second dotted configuration of compound (II) isa single bond, R₄ is H when both dotted configurations are single bonds,and R₄ is selected from the group consisting of H and OMe when bothdotted configurations are double bonds. Generally, R₅ is selected fromthe group consisting of H, OH, OMe, OR₅₀, and X wherein R₅₀ is alkyl,alkenyl, or alkynyl, and X is F, Cl, Br, or I. Preferably, R₅ isselected from the group consisting of H, OH, OMe, OR₆₀, and X whereinR₆₀ is (CH₂)_(n)C₃ and n is 1-7, ans X is F, Cl, Br, or I. Morepreferably, R₅ is selected from the group consisting of H, OH, and OMe.Even more preferably, R₅ is OH.

While compounds of formula (II) have been presented herein as diketones,and compounds of formula (I) have been presented as enols, those ofskill in the art recognize that diketones and enols can coexist insolution as tautomers as shown below.

Accordingly, the invention contemplates the use and production ofcompounds in either tautomeric form, and as a mixture of the two forms.

A natural product compound having the following general formula wasisolated from turmeric, and was found to protect cells from βApeptide-induced toxicity.

Still other turmeric-related compounds useful in practice of theinvention include those of formula (III):

In this formula, the dotted configuration is optionally a single bond ora double bond or a triple bond. Z is a representation of isostericvariation in which Z is selected from O, S, NH, NR₆₀, where R₆₀ isalkyl, alkenyl, or alkynyl. Generally, R₆ is selected from the groupconsisting of OH, OMe, OR₅₀, and X wherein R₅₀ is alkyl, alkenyl, oralkynyl, and X is F, Cl, Br, or I. Preferably, R₆ is selected from thegroup consisting of OH, OMe, OR₆₀ and X wherein R₆₀ is (CH₂)_(n)CH₃ andn is 1-7 and X is F, Cl, Br, or I. More preferably, R₆ is selected fromthe group consisting of OH and OMe. Even more preferably, R₆ is OH.Generally, R₇ is selected from the group consisting of H, OMe, and OR₅₀wherein R₅₀ is alkyl, alkenyl, or alkynyl. Preferably, R₇ is selectedfrom the group consisting of H, OMe and OR₆₀ wherein R₆₀ is (CH₂)_(n)CH₃and n is 1-7. More preferably, R₇ is selected from the group consistingof H and OMe. Even more preferably, R₇ is OMe. Generally, R₈ is selectedfrom the group consisting of OH, OMe, OR₅₀ and X wherein R₅₀ is alkyl,alkenyl, or alkynyl, and X is F, Cl, Br, or I. Preferably, R₈ isselected from the group consisting of OH, OMe, OR₆₀ and X wherein R₆₀ is(CH₂)_(n)CH₃ and n is 1-7, and X is F, Cl, Br, or I. More Preferably, R₈is selected from the group consisting of OH and OMe. Even morepreferably, R₈ is OH. Generally, R₉ is selected from the groupconsisting of H, OMe and OR₅₀ wherein R₅₆ is alkyl, alkenyl, or alkynyl.Preferably, R₉ is selected from the group consisting of H, OMe and OR₆₀wherein R₆₀ is (CH₂)_(n)CH₃ and n is 1-7. More preferably, R₉ isselected from the group consisting of H and OMe. Even more preferably,R₉ is OMe.

The second set of compounds useful for practice of the invention includenatural compounds which can be extracted on otherwise derived fromGinkgo biloba as well as synthetic Ginkgo biloba compounds includingbiologically active homologues and analogues of natural Ginkgo bilobacompounds which share anti-βA activity. Such compounds have the formula(IV):

or a pharmaceutically acceptable salt or ester thereof, wherein R isselected from the group consisting of higher alkyl, higher alkenyl, andhigher alkynyl.

More preferably, R is

and n is 1-7. Even more preferably, R is selected from the groupconsisting of

And R is also selected from the group consisting of alkyl, alkenyl, andalkynyl; for example;

and y is 1-9, or having more than one double bond (cis or trans), ortriple bond consisting of; for example;

wherein the dotted line configuration is optionally a single bond (cisor trans), or a triple bond, wherein the alkyl, alkenyl, and alkynylgroup is selected from ethers and/or thioethers or amines; for example;

wherein z=O, S, NR, where R=alkyl, alkenyl, alynyl groups; and n=1 or 2.

The third set of compounds useful for practice of the invention includenatural compounds which can be extracted on otherwise derived fromZingiber sp. (ginger) as well as synthetic ginger compounds includingbiologically active homologues and analogues of natural ginger compoundswhich share anti-βA activity. Such compounds have the formula (V):

In this formula, the dotted configuration is optionally a single bond ora double bond or a triple bond. Preferably, R₁₀ is selected from thegroup consisting of OH, OMe, OR′, and X wherein R′ is alkyl, alkenyl, oralkynyl, and X is F, Cl, Br, or I. More preferably, R₁₀ is selected fromthe group consisting of OH, OMe, OR″, and X wherein R″ is (CH₂)_(n)CH₃and n is 1-7, and X is F, Cl, Br, or I. Even more preferably, R₁₀ is OH.Preferably, R₁₁ is selected from the group consisting of H, OH, OMe, andOR′ wher R′ is alkyl, alkenyl, or alkynyl. More preferably, R₁₁ isselected from the group consisting of H, OH, OMe, and OR″ wherein R″ is(CH₂)_(n)CH₃ and n is 1-7. Even more preferably, R₁₁ is selected fromthe group consisting of H and OMe. Preferably, R₁₂ is selected from thegroup consisting of alkyl, alkenyl, and alkynyl. More preferably, R₁₂ isselected from the group consisting of

and y is 1-9.

Even more preferably, R₁₂ is selected from the group consisting of

and y is 1-9, or having more than one double bond (cis or trans), ortriple bond consisting of; for example;

wherein the dotted line configuration is optionally a single bond (cisor trans), or a triple bond, wherein the alkyl, alkenyl, and alkynylgroup is selected from ethers and/or thioethers or amines; for example;

wherein z=O, S, NR_(n), where R=alkyl, alkenyl, alynyl groups; and n=1or 2.and y is 1-9.

And compounds having a formula (VI):

In this formula, the dotted configuration is optionally a single bond ora double bond or a triple bond. Preferably, R₁₃ is selected from thegroup consisting of OH, OMe, OR′, and X wherein R′ is alkyl, alkenyl, oralkynyl, and X is F, Cl, Br, or I. More preferably, R₁₃ is selected fromthe group consisting of OH, OMe, OR″, and X wherein R″ is (CH₂)_(n)CH₃and n is 1-7, and X is F, Cl, Br, or I. Even more preferably, R₁₃ is OH.Preferably, R₁₄ is selected from the group consisting of H, OH, OMe, andOR′ wher R′ is alkyl, alkenyl, or alkynyl. More preferably, R₁₄ isselected from the group consisting of H, OH, OMe, and OR″ wherein R″ is(CH₂)_(n)CH₃ and n is 1-7. Even more preferably, R₁₄ is selected fromthe group consisting of H and OMe. Preferably, R₁₅ is selected from thegroup consisting of alkyl, alkenyl, and alkynyl. More preferably, R₁₅ isselected from the group consisting of

and y is 1-9, or having more than one double bond (cis or trans), ortriple bond consisting of; for example;

wherein the dotted line configuration is optionally a single bond (cisor trans), or a triple bond, wherein the alkyl, alkenyl, and alkynylgroup is selected from ethers and/or thioethers or amines; for example;

wherein z=O, S, NR_(n), where R=alkyl, alkenyl, alynyl groups; and n=1or 2.

It is apparent from the biological results for the ginger-derivednatural product compounds that the length of the side chain is importantfor the expression of biological activity. For example, with respect tothe ginger-derived natural product compounds, compounds (11), (12), (13)and (14), the biological activity appears to improve as the compounds'side chain length increases. Thus, it is of interest to prepareanalogues having different and lengthier side-chains. Preferably,shogaol compounds have side chains wherein R₁₂ has five or more carbons.More preferably, R₁₂ has nine or more carbons, and even more preferably,R₁₂ has eleven or more carbons. Furthermore, two of the synthesizedshogaol analogue compounds, compounds (45) and (50), also effectivelyprotected cells from PA peptide insult despite the fact that thesecompounds have different substituents than the ginger-derived naturalproduct compounds. For example, compound (45) differs from theginger-derived natural product compounds because it has a saturatedhydrocarbon side chain, and compound (50) differs from theginger-derived natural product compounds because it does not have amethoxy substituent. These data suggest that changing the nature of thesubstituents on the phenyl rings of the active compounds is of interestfor the methods, pharmaceutical compositions, compounds and usesaccording to the invention.

As used herein, the term “alkyl” refers to a carbon chain having atleast two carbons. Preferably, alkyl refers to a carbon chain havingbetween two and twenty carbons. More preferably, alkyl refers to acarbon chain having between two and eight carbons. The term “alkenyl,”as used herein, refers to a carbon chain having at least two carbons,and at least one carbon-carbon double bond. Preferably, alkenyl refersto a carbon chain having between two and twenty carbons, and at leastone carbon-carbon double bond. More preferably, the term alkenyl refersto a carbon chain having between two and eight carbons, and at least onecarbon-carbon double bond. The term “alkynyl,” as used herein, refers toa carbon chain having at least two carbon atoms, and at least onecarbon-carbon triple bond. Preferably, alkynyl refers to a carbon chainhaving between two and twenty carbon atoms, and at least onecarbon-carbon triple bond. More preferably, alkynyl refers to a carbonchain having between two and eight carbon atoms, and at least onecarbon-carbon triple bond.

As used herein, the term “higher alkyl” refers to a carbon chain havingat least five carbon atoms. Preferably, higher alkyl refers to a carbonchain having between five and twenty carbons. More preferably, higheralkyl refers to a carbon chain having between five and twelve carbonatoms. As used herein, the term “higher alkenyl” refers to a carbonchain having at least five carbon atoms, and at least one cabon-carbondouble bond. Preferably, higher alkenyl refers to a carbon chain havingbetween five and twenty carbon atoms, and at least one carbon-carbondouble bond. More preferably, higher alkenyl refers to a carbon chainhaving between five and twelve carbon atoms, and at least onecarbon-carbon double bond. The term “higher alkynyl,” as used herein,refers to a carbon chain having at least five carbons, and at least onecarbon-carbon triple bond. Preferably, higher alkynyl refers to a carbonchain having between five and twenty carbon atoms, and at least onecarbon-carbon triple bond. More preferably, the term higher alkynylrefers to a carbon chain having between five and twelve carbon atoms,and at least one carbon-carbon triple bond.

The fourth set of compounds useful for practice of the invention includenatural compounds which can be extracted or otherwise derived fromSalvia sp. (sage) and Rosmarinus sp. (rosemary) which share anti-βAactivity. Such compounds have the formulas (VII), (VIII) and (IX):

The administration of the natural product and natural product analoguecompounds of the invention is preferably accomplished with apharmaceutical composition comprising a therapeutically effective amountof an active compound of the present invention and a pharmaceuticallyacceptable diluent, adjuvant, or carrier. A compound according to theinvention may be administered without or in conjunction with knownantibiotics, surfactants, or other therapeutic agents. It iscontemplated that the pharmaceutical compositions of this invention canbe administered to humans and other animals orally, rectally,parentally, intracisternally, intraperitoneally, intraocularly byinjection or depot, topically (as by powders, ointments, or drops),intraocularly, bucally, intranasally, or by any other effective route ofadministration.

According to the methods for treatment of the present invention, βAprotein-induced disease is treated in a subject, such as a human orlower mammal, by administering to the subject a therapeuticallyeffective amount of an active compound of the invention in such amountsand for such time as is necessary to achieve the desired results. Theterm “beta-Amyloid protein-induced disease”, as used herein, refers todisease states that are characterized by the formation and aggregationof beta-Amyloid protein or beta-Amyloid peptide fibril deposits orplaques, such as, for example, Alzheimer's disease, Down's syndrome,age-related macular degeneration (AMD) and glaucoma.

It is contemplated that the methods for treatment in accordance with theinvention encompass the treatment of subjects wherein the βAprotein-induced disease process is ongoing but wherein the subjects donot exhibit manifest outward symptoms, and/or wherein the pathology ofthe disease can not be detected using presently available technologies.Furthermore, the methods for treatment of the present inventioncontemplate not only treating the common symptoms associated with βAprotein-induced diseases but also treating the pathology of the disease.Thus, the methods for treatment provided herein include treatingsymptoms associated with βA protein-induced diseases, such as, forexample, the memory loss and dementia associated with Alzheimer'sdisease, but also include preventing senile plaque formations, and/orclearing such formations. Similarly, the methods of the invention arecontemplated to be useful in treating the symptoms associated withβA-induced ocular diseases such as glaucoma and AMD and further treatthe pathology of those diseases. It is hypothesized that the formationof senile plaques is a regularly occurring and ongoing process in humansand other mammals. However, it is further hypothesized that theequilibrium of this process is substantially disturbed in patientsaffected by βA protein-induced diseases, resulting in the accumulationand formation of senile and ocular plaques.

As used herein, the term “therapeutically effective amount” means thatamounts of a compound of the present invention sufficient to alleviate,ameliorate, prevent, and/or clear the symptoms and/or the pathology ofβA protein-induced disease are contemplated for administration.Accordingly, the methods for treatment of AD in accordance with theinvention contemplate administration of an active compound of theinvention whether βA protein-induced disease-like symptoms are manifest,or not.

The total daily dose of natural product compound (6) of this inventionto be administered to a human or other mammal is preferably between 1 to200 mg/kg body weight. More preferably, the total daily dosage isbetween 20 to 160 mg/kg body weight. Even more preferably, the totaldaily dosage is between 40 to 100 mg/kg body weight. One skilled in theart could obtain preferred dosage ranges for the other compounds of theinvention by extrapolating from the compounds' ED₅₀ values, such as, forexample the ED₅₀ values presented in Tables 1, 2, 3, and 4. It will beunderstood that the total daily usage of the compounds and compositionsof the present invention will be decided by the attending physicianwithin the scope of sound medical judgment. The specific therapeuticallyeffective dose level for any particular patient will depend upon avariety of factors including the severity and progression of thedisease, the time of administration, the route of administration, thesize of the subject, the rate of excretion of the specific compoundemployed, the duration of the treatment, the additional therapeuticagents used in combination with the specific compound of the invention,and like factors well known in the medical arts.

The mechanism of action of the natural product compounds and the naturalproduct analogue compounds of the invention appears to involve bothantioxidant and non-antioxidant pathways. Without intending to be boundby a theory of the mechanism of the invention, it is believed that thecompounds and compositions of the invention provide therapeutic andpreventive agents that protect neurons from βA peptide insult by (1) anantioxidant pathway, (2) preventing the aggregation of βA peptide bydirectly binding to PA peptide, thereby altering its structuralconformation and rendering it non-toxic, and/or (3) binding to areceptor site on the cell, thereby altering the cell function in such away that it is protected from βA peptide toxicity.

The invention can be better understood in light of the followingexamples which are intended as an illustration of the practice of theinvention and are not meant to limit the scope of the invention in anyway.

EXAMPLE 1 Isolation and Identification of Natural Product CompoundsDerived from Turmeric that Protect Cells from Beta Amyloid-InducedToxicity

According to this example, potent anti-AD natural product compounds thatprotect cells from βA peptide-induced toxicity were isolated fromturmeric by following bioassay-guided fractionation schemes. Briefly,ground turmeric was extracted with 90% methanol overnight (2×), and thesolvent was removed under vacuum at 35° C. The residue was partitionedbetween petroleum ether/water, dichloromethane/water, and ethylacetate/water, successively. After removing the solvents under vacuum at35° C., the residues from each partition were screened for inhibitoryactivity against βA peptide-induced cytotoxicity using the MTT assaydescribed in this example. The active principles were isolated from theresidues of the active fractions by a series of column chromatographyusing various resins (Amberchrom non-ionic resin and silica gel) andsemi-preparative HPLC reverse-phased separation (isopropyl alcohol/wateror acetonitrile/water solvent system). Six curcuminoids, natural productcompounds (1), (2), (3), (4), (5), and (6) were isolated from turmeric,and their structures were elucidated using NMR (1-D and 2-D ¹H, ¹³C,APT, HMBC) and mass spectrum analysis. These compounds are shown in FIG.1.

The inhibitory activity of the residues and of the identified compoundswas determined by observing the differences in the cell viability of βApeptide (both 25-35 and 1-42) treated cells, βA peptide (both 25-35 and1-42) treated cells further including a compound according to theinvention, and a DMSO control.

The degree of βA insult was measured by3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT)reduction assay. See Kim et al., Neurosci Lett 303, 57 (2001), Park etal., J Nat Prod 65, 1227 (2002) and Kim et al., Plant Medica 68, 375(2002). The detection of cell growth or cell death can be determined byobserving the conversion of MTT to the colored product, MTT formazan,the concentration of which can be measured calorimetrically at 550 nm.See Kim et al., Neurosci Lett 303,57 (2001).

The βA peptide-induced toxicity inhibitory effects of the compounds weretested on PC12 cells. The cells were incubated with βA peptide (25-35)(1.0 μg/ml, made from 1.0 mg/ml stock solution in DMSO) or βA peptide(1-42) (2.0 μg/ml, made from 1.0 mg/ml stock solution in DMSO) and thetest compounds at various concentrations (25, 5.0, 1.0, and 0.2 μg/ml)in collagen-coated 96-well tissue culture plates for 24 hours. The PApeptide-induced toxicity inhibitory effect of the compounds wasdetermined by calorimetrically and microscopically evaluating the PC12cells' potential to reduce MTT against a positive control (1% DMSO only)and a negative control (1.0 μg/ml βA peptide in 1% DMSO alone). Cellswere incubated in MTT solution (5 mg/ml) at 37° C. for 2 hours. Duringthis time, cells were observed under a microscope every 15 min. Cellswere incubated in Lysing buffer (100 μl) overnight at 37° C.Colorimetric determination of MTT reduction was made at 550 nm. The βApeptide-induced cytotoxicity inhibitory activity of the compounds wasalso evaluated against IMR32, HUVEC, and primary cortical rat neuronalcells.

PC12 rat pheochromocytoma and IMR32 human neuroblastoma cells wereobtained from the American Type Culture Collection (ATCC). HUVEC normalumbilical human vein endothelial cells were obtained from Clonetics (SanDiego, Calif.). Cells were routinely cultured on a tissue culture plate(Corning, N.Y., N.Y.). PC12 cells were grown in high glucose Dulbecco'sModified Eagle Medium (DMEM), 10% horse serum, 5% fetal calf serum, and1% penicillin/streptomycin. IMR32 cells were grown in 90% DMEM and 10%fetal calf serum with 1% penicillin/streptomycin. HUVEC cells were grownin EGM-2 Bullet Kit (Clonetics, San Diego, Calif.). For the bioassayusing βA peptide(25-35) and βA peptide(1-42), 100 μl of exponentiallygrowing PC12 cells (2,000 cells per ml) were plated in collagen-coated96-well tissue culture plates.

PC12 cells were cultured routinely on polystyrene-coated Corning tissueculture plates. PC12 cells gave consistent results only when thecollagen-coated 96-well plates were used. The 96-well plates were coatedwith rat tail collagen (Boehringer Mannheim) in order to promote uniformPC12 cell attachment and growth. Under the experimental conditions, βApeptide(25-35) and βA peptide(1-42) was toxic to PC12 cells at ED₅₀=1.0and 5.0 μg/ml, respectively.

IMR32 and HUVEC cells were chosen to confirm and supplement the anti-PApeptide activity of the compounds identified by the assay using PC12cells. βA peptide has been reported to be cytotoxic to IMR32 andendothelial cells. Experimental results demonstrated that IMR32 andHUVEC cells are sensitive to βA peptide (25-35) at ED₅₀=3.0 and 6.0μg/ml, respectively, and βA peptide (1-42) at ED₅₀=6.0 and 10.0 μg/ml,respectively.

Primary cortical rat neuronal cells were obtained in the followingmanner. Dissociated primary neuronal cell cultures were established from18-day-old Sprague-Dawley rat fetuses. The pups were delivered bycaesarean section while the dam was anesthetized with ether. Hippocampaltissue from embryonic day 18 Sprague-Dawley rat pups was dissected andthen rinsed in cold Ca²⁺/Mg²⁺-free Hank's balanced salt solutionsupplemented with 20 mM HEPES, 4.2 mM sodium bicarbonate, 1 mM pyruvate,and 3 mg/ml bovine serum albumin (BSA). Following gentle trituration ofthe tissue with a constricted pipette in cold buffer, two volumes of 10%fetal bovine serum (FBS) in DMEM were added to the suspension. After thesuspension settled for 2 minutes, the supernatant was collected andcentrifuged for 2 min. at 200×g. The cell pellets were resuspended inserum-free DMEM (pH 7.3), supplemented with 2.4 mg/ml BSA and amodification of Brewer's B16 defined components (with 250 nM vitamin B12and without catalase, glutathione, and superoxide dismutase). Cells wereplated at a density of 15,000 cells/cm² and grown at 37° C. After 24hours of incubation to allow cell attachment, the serum-containingmedium was replaced by defined medium with DMEM/F12 containing bovinetransferrin (100 μg/ml), bovine insulin (5 μg/ml), putrescine (0.1 mM),progesterone (10 nM), sodium selenite (30 nM), sodium pyruvate (1 mM),and potassium bicarbonate (15 mM). Cells maintained for extended periodsof time were fed twice a week by replacing ⅓ of the medium with freshmedium.

EXAMPLE 2 Inhibitory Activity and Antioxidant Potency ofTurmeric-Derived Natural Product Compounds Against Beta Amyloid Toxicity

According to this example, the inhibitory activity of theturmeric-derived natural product compounds (1), (2), (3), (4), (5), and(6) (shown in FIG. 1) against PA peptide-induced toxicity was measuredby the MTT reduction assay described in example 1. These sixturmeric-derived curcuminoids protected PC12, IMR32, and HUVEC cellsfrom βA peptide-induced toxicity (Table 1). These compounds alsoprotected primary cortical neuronal cells at 5 μg/ml against βApeptide(1-42) insult (10 μg/ml).

ED₅₀ values reflect the results from the MTT assay, and represent thesample concentration that is required to achieve 50% cell viability, amid-point between the positive control values and the negative controlvalues. The samples that gave values as determined by the MTT assay lessthan or equal to that of βA peptide treated wells were consideredcytotoxic or without desired activity, and are labeled “toxic”.

The measurement of lactate dehydrogenase activity released to theextracellular bathing media was also used to assess cell viability incell culture. LDH activity in the medium was measured. See Kimura etal., Brain Res 1047, 72 (2005) and Loudina et al., Exp Neurol 184, 923(2003). This assay was used to confirm the ED₅₀ results obtained in theMTT assay. Samples of media from 96-well cell culture plates weretransferred to an empty well of a 96-well plate (100 μl) and 2.0 μmol ofsodium pyruvate and 0.1 mg of the reduced form of nicotinamide adeninedinucleotide (NADH in 0.1 M K₂PO₄ buffer (pH 7.5 at 25° C.) were added(total volume of 400 μl). The absorbance of the reaction mixture at 340nm provides an index of NADH concentration, and was recorded using aspectrophotometer 5 minutes after mixing the reagents. The experimentwas performed in triplicate and the LDH concentration was calculatedfrom the slope of the absorbance curve, fit by linear regression to thelinear (initial) portion of the curve. The concentration of LDH wasexpressed in conventional units (u) per ml. Accuracy of the assay wasverified by periodic checks of a standard LDH enzyme solution (Sigma).

IC₅₀ values reflect the results of the antioxidant assay described inthis example, and represent the sample concentration which is requiredto scavenge 50% of the DPPH free radicals. Kim et al., Neurosci Lett303, 57 (2001) and Barik et al., Free Radic Biol Med 39, 811 (2005).

Using an antioxidant assay, the antioxidant potency of the compounds ofthe invention was evaluated. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) isknown to generate stable free radicals in aqueous and ethanolicsolutions. The ability of the compounds of the invention to scavengethese free radicals was measured by observing the optical density changeof DPPH radicals at 515 nm. Kim et al., Neurosci Lett 303, 57 (2001) andBarik et al., Free Radic Biol Med 39, 811 (2005).

The samples were prepared in various concentrations (200, 20, 2.0, and0.2 μg/ml) by serial dilution of a stock solution (5 mg/ml) and weretested by the following procedure. Reaction mixtures containing testcompounds (dissolved in DMSO) and 300 μM DPPH ethanolic solution in96-well microtiter plates were incubated at 37° C. for 30 min. andabsorbance was measured at 515 nm. Percent inhibition by sampletreatment was determined by comparison with a DMSO-treated positivecontrol group. IC₅₀ values were determined from percent inhibition bysample. IC₅₀ values denote the concentration of the tested compound thatwas required to scavenge 50% of the DPPH free radicals.

The antioxidant potency of the natural product compounds was evaluatedby measuring the compounds' ability to scavenge free radicals in orderto elucidate the possible involvement of antioxidant pathways in thecompounds ability to protect the cells (Tables 1 and 2). The resultsshowed that only compounds (1) and (2) have strong antioxidant activity,suggesting that the compounds of the invention may be protecting cellsfrom βA peptide insults through a mechanism that does not involve anantioxidant pathway. TABLE 1 Inhibitory Activity of Turmeric-DerivedNatural Product Compounds against βA Peptide-Induced Toxicity againstPC12, IMR32, and HUVEC cells and Antioxidant Activity of the Compounds.Anti-β A Anti-βA Anti-βA Anti-βA Anti-βA peptide(25-35) peptide(1-42)peptide(25-35) peptide(25-35) peptide(1-42) ED₅₀ (μg/ml) ED₅₀ (μg/ml)ED₅₀ (μg/ml) ED₅₀ (μg/ml) ED₅₀ (μg/ml) Antioxidant Compound PC12 PC12IMR32 HUVEC HUVEC IC₅₀ (μg/ml) 1 7.0 10 6.0 12 13 28.2 2 4.0 5.0 4.0 4.55.0 36.2 3 2.0 3.5 2.5 2.4 2.0 >200 4 0.5 1.0 1.2 0.8 1.0 >200 5 2.5 3.01.5 2.0 1.5 >200 6 1.0 2.0 1.0 1.5 1.0 >200

EXAMPLE 3 Curcuminoid Analogue Synthesis

According to this example, curcuminoids and curcuminoid analogues weresynthesized.

Dihydro- and tetrahydro-curcuminoids were synthesized by the procedureillustrated in FIG. 2. 3-(4-hydroxyphenyl) propanoic acid, compound(18), was treated with TMSCl (1.3 equivalents) in the presence of 1.1equivalents of triethylamine in THF/CH₂Cl₂ (50/50) solution toprecipitate triethylammonium chloride as a white solid. The reaction wasover within a few minutes, and only the phenolic position was protected.The white ammonium salt was filtered, and the filtrate was diluted withethyl acetate. The resulting solution was washed with water three times,dried (MgSO₄), filtered, and the solvent was removed under vacuum toafford the TMS protected carboxylic acid, compound (19), in quantitativeyield. The TMS protected carboxylic acid, compound (19), was convertedto the corresponding acyl chloride, compound (20), by refluxing inoxalyl chloride for 30 min, and the remaining oxalyl chloride wasremoved under a stream of N₂ gas. 4-(4-hydroxyphenyl)-2-butanone,compound (21), was treated with TMSCl (1.3 equivalents) in the presenceof triethylamine (1.1 equiv) in CH₂Cl₂, yielding the TMS protectedproduct, compound (22). The ammonium chloride precipitate was filtered,and the filtrate was diluted with ethyl acetate. The resulting solutionwas washed with water (3×), dried (MgSO₄), filtered, and the solvent wasremoved under vacuum to afford compound (22), in quantitative yield.Compound (22) was treated with lithium diisopropylamide (LDA, 1.5 M inTHF, 1. equiv) in tetrahydrofuran (THF) at −78° C. under N₂ for 20 minand 1.1 equivalents of the TMS protected acyl chloride, compound (20),dissolved in THF was added. The reaction mixture was stirred at −78° C.for 15 minutes and slowly warmed to room temperature. The reactionmixture was quenched with water and poured into ethyl acetate. Theorganic layer was washed three times with water and the water layer wasback washed (2×) with ethyl acetate. The organic layers were combined,dried (MgSO₄), filtered, and the solvent was removed under vacuum. Theresidue was stirred in methanol in the presence of K₂CO₃ for 30 min toremove TMS protection. The solution was acidified with 2N HCl and pouredinto ethyl acetate. The aqueous layer was partitioned three times withethyl acetate and the organic layers were combined, dried (MgSO₄),filtered, and the solvent was removed under vacuum. The residue wascolumn chromatographed over silica gel using a gradient elution of ethylacetate/petroleum ether to afford compound (9). Compound (9) was furtherpurified using semi-preparative HPLC using an acetonitrile/water (90/10)solvent system to give pure synthesized curcuminoid compound (9) in 45%overall yield. Similarly, the unsymmetric synthesized curcuminoidcompound (4) was prepared in 40% overall yield.

Both symmetric and unsymmetric curcumin analogues and related compoundswere prepared according to the procedure described in FIG. 3.Benzaldehyde, compound (24), 4-hydroxybenzaldehyde, compound (25),2,4-pentadione, compound (26), and boric acid were dissolved in dryN,N-dimethylformamide (DMF), and treated with a small amount of1,2,3,4-tetrahydroquinoline and glacial acetic acid. This reactionyielded three products: a diphenyl group substituted product, compound(27), in 31% yield, a dihydroxyphenyl group substituted product,compound (3), in 6% yield, and a hydroxyphenyl phenyl substitutedproduct, compound (28), in 11% yield. After working up the reaction,(ethyl acetate/water partitioning and back washing of the aqueous layerwith ethyl acetate, followed by drying (MgSO₄) of the organic layer andremoval of solvent in vacuo), the products were separated usingsemi-preparative HPLC (75% isopropyl alcohol/H₂O eluent system). Thephysical data (¹H NMR) of the dihydroxyphenyl product (3) was identicalto that of the turmeric-derived natural product (3).

Natural product compound (6) was synthetically prepared according to theprocedure shown in FIG. 4. The alcohol functionalities of acetol,compound (29), and vanillin, compound (31), were protected inquantitative yield as tetrahydropyran (THP) ethers using dihydropyran(DHP) in the presence of pyridinium para-toluene sulfonate (PPTS) inTHF. The THP ether of acetol, compound (30), was reacted with LDA in THFat −78° C. and then reacted with the THP ether of vanillin, compound(32), to afford the β-hydroxy ketone, compound (33), in 73% yield. TheTHP ether was removed in the presence of PPTS, causing the dehydrationof the β-hydroxyl group, and affording compound (34) in 72% yield. Thephenolic group of compound (34) was selectively protected with a TMSgroup in quantitative yield to yield an alcohol, compound (35). Thephenolic group of 4-hydroxy-3-methoxyphenyl propenoic acid, compound(36), was selectively protected with a TMS group in quantitative yield.The TMS protected carboxylic acid, compound (37), and the alcohol,compound (35), were coupled in the presence of dicyclohexylcarbodiimide(DCC) and dimethylamino-pyridine (DMAP) in THF at room temperature toafford 68% of the coupled product, compound (38). The TMS protectinggroups of compound (38) were removed by stirring in a mixture of aceticacid/H₂O in THF (1/1/5) to afford the desired product in 53% yield.Attempts to remove the TMS groups of compound (38) usingtetra-n-butylammonium fluoride in THF resulted in the decomposition ofthe desired reaction product. The ¹H NMR of the product was identical tothat of turmeric-derived natural product compound (6).

EXAMPLE 4 Inhibitory Activity and Antioxidant Potency of CurcuminoidSynthetic Analogues Against Beta Amyloid-Induced Toxicity

According to this example, the inhibitory activity of the syntheticcurcuminoid analogues against βA peptide-induced toxicity was measuredby the MTT reduction assay described in example 1. Synthesized compounds(1), (3), (4), and (9) (shown in FIG. 5) protected the cells from βApeptide insult (Table 2). Microscopic analyses of βA peptide treatedcells further including synthesized curcuminoid compounds (3) and (4)also demonstrated that these compounds effectively protect cells from βApeptide insults. The positive control and cells treated with compounds(3) and (4) maintained MTT formazan granules in the cytosole, a sign ofviable cells, while the negative control showed extensive MTT formazanspike processes, a sign of nonviable cells. As was the case with thestructurally analogous natural product compound, natural productcompound (4), synthesized curcuminoid compound (4) provided the bestprotection. Interestingly, synthesized curcuminoid compounds (7), (8),and (10) were cytotoxic. Apparently, the presence of a hydroxyl group atthe 4-position of phenyl ring or the size of substituent at thatposition is important for the expression of the desired biologicalactivity. The results of the MTT assay were confirmed by the LDHmethodology set forth in example 2. The synthesized curcuminoidcompounds are shown in FIG. 5.

The ability of the synthesized curcuminoid compounds to scavenge DPPHfree radicals was measured by observing the optical density change ofthe radicals at 515 nm in accordance with the antioxidant assay setforth in Example 2. The results show that only compounds 1 and 3 havesignificant antioxidant activity (Table 2). TABLE 2 Inhibitory Activityof Synthesized Curcuminoids against βA Peptide-Induced Toxicity againstPC12 and IMR32 Cells and Antioxidant Activity of the Compounds. Anti-βAAnti-βA Anti-βA Anti-βA peptide(25-35) peptide(1-42) peptide(25-35)peptide(1-42) ED₅₀ (μg/ml) ED₅₀ (μg/ml) ED₅₀ (μg/ml) ED₅₀ (μg/ml)Antioxidant Compound PC12 PC12 IMR32 IMR32 IC₅₀ (μg/ml) 1 5.5 6.0 6.06.0 28.5 3 3.0 4.5 3.0 3.5 32.6 4 0.5 1.0 1.5 2.0 >200 7 toxic toxictoxic toxic >200 8 toxic toxic toxic toxic >200 9 10.0 9.0 12.011.0 >200 10 toxic toxic toxic toxic >200

EXAMPLE 5 Isolation and Identification of Natural Product CompoundsDerived from Ginger that Protect Cells from Beta Amyloid-InducedToxicity

According to this example, natural product compounds that protect cellsfrom βA peptide-induced toxicity were isolated from ginger by followingbioassay-guided fractionation schemes. Briefly, ground ginger wasextracted with 90% methanol overnight (2×), and the solvent was removedunder vacuum at 35° C. The residue was partitioned between petroleumether/water, dichloromethane/water, and ethyl acetate/water,successively. After removing the solvent under vacuum at 35° C., theresidues from each partition were screened for inhibitory activityagainst βA peptide-induced cytotoxicity using PC12, IMR32, and HUVECcells at 25, 5.0, and 1.0 βg/ml. The active principles were isolatedfrom the residues of active fractions by a series of columnchromatography using various resins (Amberchrom non-ionic resin andsilica gel) and semi-preparative HPLC reverse-phased separation(isopropyl alcohol/water or acetonitrile/water solvent system). Fourshogaols, natural product compounds (11), (12), (13), and (14) (shown inFIG. 6) were isolated from ginger, and their structures were elucidatedusing NMR (1-D and 2-D ¹H, ¹³C, APT, HMBC) and mass spectrum analysis.

EXAMPLE 6 Inhibitory Activity of Ginger-Derived Natural ProductCompounds Against Beta Amyloid Toxicity

According to this example, the inhibitory activity of natural productcompounds (11), (12), (13), and (14) (shown in FIG. 6) against βApeptide-induced toxicity was measured by the MTT reduction assay setforth in example 1. These natural product compounds effectivelyprotected PC 12, IMR32, and HUVEC cells from βA peptide-induced toxicity(Table 2). The results of the MTT assay were confirmed by the LDHmethodology set forth in example 2.

The ability of natural product compounds (11), (12), (13), and (14) toscavenge DPPH free radicals was measured by observing the opticaldensity change of the radicals at 515 nm in accordance with theantioxidant assay set forth in Example 2. None of these compoundsexhibited significant antioxidant activity. TABLE 3 Inhibitory Activityof Ginger-Derived Natural Product Compounds against βA Peptide-InducedToxicity against PC12, IMR32, and HUVEC cells and Antioxidant Activityof the Compounds. Anti-βA Anti-βA Anti-βA Anti-βA Anti-βA peptide(25-35)peptide(1-42) peptide(25-35) peptide(25-35) peptide(1-42) ED₅₀ (μg/ml)ED₅₀ (μg/ml) ED₅₀ (μg/ml) ED₅₀ (μg/ml) ED₅₀ (μg/ml) Antioxidant CompoundPC12 PC12 IMR32 HUVEC HUVEC IC₅₀ (βg/ml) 11 15 12 15 20 20 >200 12 9.010 8.0 20 18 >200 13 3.0 4.0 2.0 8.0 8.0 >200 14 2.0 2.0 1.5 4.0 5.0>200

EXAMPLE 7 Shogaol Analogue Synthesis

According to this example, shogaols and their analogues weresuccessfully synthesized in 100 mg scale. Gingerols were synthesizedfrom zingerone by conversion into the corresponding O-trimethylsilylether, deprotonation with lithium bis(trimethylsilyl)amide or lithiumdiisopropylamide (LDA), and regioselective aldol condensation. Shogaolsare gingerol analogues with a 4,5-double bond, resulting from theelimination of the 5-hydroxy group.

The phenol group of vaniline, compound (31), was protected as the THPether (DHP/PPTS/CH₂Cl₂) to yield compound (32), and the aldehyde groupof compound (32) was reduced to the alcohol to yield compound (39) usingNaBH₄ in THF as shown in FIG. 7. The resulting alcohol, compound (39),was mesylated (methanesulfonyl chloride/triethylamine/THF) and thenreacted with in situ generated lithium acetonide (acetone/LDA/THF/−78°C.) at −78° C. under N₂ to yield compound (41). Compound (41) wasreacted with LDA at −78° C. in THF under N₂ to generate lithium enolatewhich was then reacted with octyl aldehyde to afford the β-hydroxyketone, compound (42). During treatment with PPTS in ethanol at 50° C.to remove the THP ether protecting group, dehydration occurred to afford[9]-shogaol, compound (13), which was identical to the ginger-derivednatural product compound (13) (overall yield 37%).

The phenol group of vaniline, compound (31), was protected as the THPether (DHP/PPTS/CH₂Cl₂) to yield compound (32), and the aldehyde groupof compound (32) was reduced to the alcohol to yield compound (39) usingNaBH₄ in THF as shown in FIG. 8. The resulting alcohol, compound (39),was mesylated (methanesulfonyl chloride/triethylamine/THF) and reactedwith in situ generated lithium 2-undecanonide, compound (44),(2-undecanone/LDA/THF/−78° C.) at −78° C. under N₂. The THP etherprotecting group was removed by further treating the reaction mixturewith PPTS in ethanol at 50° C. to afford [9]-dihydroshogaol, compound(45).

The phenol group on 4-(4-hydroxy-phenyl)-2-butanone, compound (46), wasprotected as the TMS ether (TMSCl/triethylamine/THF) at room temperatureas shown in FIG. 9. The resulting ketone, compound (47), was reactedwith LDA at −78° C. in THF under N₂ to generate lithium enolate whichwas reacted with octyl aldehyde compound (48), to afford the β-hydroxyketone, compound (49). The TMS group was removed by stirring with NaHCO₃in methanol at room temperature. Dehydration of the β-hydroxy group wasachieved by further treatment of the reaction mixture with methanolicHCl (1 N) at room temperature to afford [9]-demethoxyshogaol, compound(50).

EXAMPLE 8 Isolation and Identification of Natural Product CompoundsDerived from Gingko Biloba that Protect Cells from Beta Amyloid-InducedToxicity

According to this example, freshly ground fresh ginkgo nuts (1 kg) wereextracted with methanol (2×2000 ml) and sequentially partitioned withpetroleum ether, ethyl acetate, dichloromethane, and butanol. Thepetroleum ether and ethyl acetate fractions protected PC12 and HUVECcells from βA peptide(25-35)-induced cytotoxicity at ED₅₀=10 μg/ml. Theactive principles were isolated from the residues of active fractions bya series of column chromatography using various resins (Amberchromnon-ionic resin and silica gel) and semi-preparative HPLC reverse-phasedseparation (isopropyl alcohol/water or acetonitrile/water solventsystem). The structures of the compounds were elucidated using 1-D and2-D NMR techniques that include ¹H, ¹³C, HMBC, and APT. Cis conformationof the double bond was unambiguously assigned in the ¹H NMR spectrum.The position of the double bond was elucidated by oxidatively cleavingit to acid functionality (KMnO₄ oxidation) and observing the massspectral fragmentation pattern (EI 70 eV). The two compounds (15) and(16) have ginkgolic acid structures, and are shown in FIG. 10. Thesecompounds have been previously isolated from ginkgo leaves, See Jaggy etal., Pharmazie 52, 735 (1997).

EXAMPLE 9 Inhibitory Activity of Ginkgo Biloba-Derived Natural ProductCompounds Against Beta Amyloid Toxicity

According to this example, the inhibitory activity of natural productcompounds derived from ginkgo biloba against βA peptide-induced toxicitywas measured by MTT reduction assay. The two ginkgo biloba-derivednatural product compounds that do not possess antioxidant properties,compounds (15) and (16), were found to protect PC12, IMR32, and HUVECcells from βA peptide-induced toxicity. The results of the MTT assaywere confirmed by following the LDH methodology set forth in example 2.This example also provides data indicating that the ginkgolides A, B,and C, (−)bilobalide, and quercetin do not possess biological activityagainst βA peptide as had been postulated in the prior art. TABLE 4Inhibitory Activity of Ginkgolic Acids 1 and 2, Ginkgolide A, GinkgolideB, Ginkgolide C, (−)- Bilobalide, and Quercetin Toward β- Insult AgainstPC12, IMR32, and HUVEC Cells. Anti-βA Anti-βA Anti-βA Anti-βA Anti-βAAnti-βA peptide(25-35) peptide(1-42) peptide(25-35) peptide(1-42)peptide(25-35) peptide(1-42) ED₅₀ (μg/ml) ED₅₀ (μg/ml) ED₅₀ (μg/ml) ED₅₀(μg/ml) ED₅₀ (μg/ml) ED₅₀ (μg/ml) Compound PC12 PC12 IMR32 IMR32 HUVECHUVEC 15 3.0 2.0 3.5 2.5 5.0 1.5 16 2.0 1.0 2.0 1.0 2.5 1.0 Ginkgolide Atoxic toxic toxic toxic toxic toxic Ginkgolide B toxic toxic toxic toxictoxic toxic Ginkgolide C toxic toxic toxic toxic toxic toxic(−)-Bilobalide toxic toxic toxic toxic toxic toxicQuercetin >20 >20 >20 >20 >20 >20

EXAMPLE 10 A Proposed Gingkolic Acid Synthesis

According to this example, a gingkolic acid synthesis is proposed asshown in FIG. 11. The benzoic acid, compound (60) and an alkyne having aterminal carbon-carbon triple bond, R, are treated withtetrakis(triphenylphosphine)palladium, in the presence of diisopropylamine and copper(I) iodide to yield an alkyne substituted benzoic acid,compound (61). Compound (61) is treated with LDA in THF, the temperatureis lowered to −78° C. and the reaction mixture is treated withoxodiperoxymolybdenum (pyridine)-(hexamethylphosphoric triamide) (MoOPh)to yield a hydroxy functionalized product, compound (62). Compound (62)is then reacted with hydrogen gas over a palladium/carbon catalyst, andtreated with acetic acid to yield the desired ginkgolic acid product,compound (63).

EXAMPLE 11 Control Study

As a control study, vitamin A, β-carotene, vitamin C, and vitamin E weretested for both anti-βA peptide(25-35) and anti-βA peptide(1-42)activity. Since these vitamins are suggested for the delaying the onsetof AD, the biological activity of the compounds of the invention werecompared with these vitamins. Under the experimental conditions, thesevitamins did not protect PC12 cells from βA peptide insults even at 200μg/ml. Congo red was also tested because it has been reported to inhibitβA peptide fibril-induced toxicity against PC 12 cells. At highconcentrations of Congo red (>25 μg/ml), the data from the cellviability evaluation using MTT reduction assay was not reliable becauseof the dye's intense red color. Nevertheless, the natural productcompounds (1), (2), (3), and (4) and natural product compounds (11),(12), (13), and (14) (ED₅₀=20.0-0.5 μg/ml) are more than 20-40 times aseffective in protecting PC 12 cells against βA peptide insults whencompared with these vitamins and other agents.

EXAMPLE 12 Reduced Glutathione Assisted βA Peptide Toxicity InhibitionAssay

According to this example, the compounds of the invention were evaluatedto ascertain if their antioxidant potency was increased whenadministered in conjunction with reduced glutathione. The synergisticinteraction between estrogens and the intracellular antioxidant, reducedglutathione (GSH), was reported to protect neurons from βApeptide-induced toxicity. See Barkats et al., J Neurochem 75, 1438(2000) and Muller et al., J Neurochem 68, 2371 (1997). The possibleinvolvement of this mechanism was evaluated using PC12 cells with thecompounds of the invention. The dose of GSH used in this study wascomparable to the low micromolar GSH (3.25 μM) concentrations found inthe cerebrospinal fluid and used by Green et al. It was hypothesizedthat if the compounds' of the invention ability to protect cells from βApeptide-induced toxicity resulted from the compounds' antioxidantpotency, administration of a compound of the invention concurrently withGSH should improve the ED₅₀ and IC₅₀ values for the compounds. Under theexperimental conditions, GSH did not influence the compounds' ability toprotect cells from βA peptide insults, and did not enhance theantioxidant potency of the compounds.

EXAMPLE 13 Determination of Ability of Compounds of the Invention toPass Through the Blood Brain Barrier

According to this example, the ability of the compounds of the inventionto pass through the blood brain barrier was measured. The ability ofcompounds to cross the blood brain barrier is represented by the log ofthe partition coefficient (P) of a molecule of the invention betweenwater and octane alcohol. Natural product compounds (1) and (3) werefound to have log P values of 3.4 and 3.1, respectively. Accordingly,the octane alcohol fraction contained more than 1000 times as much ofthe compounds as the water fraction. These results suggest that thecompounds are able to cross the blood brain barrier. See Hau et al.,Regul Toxicol Pharmacol 35, 273 (2002) and Salminen et al., J PharmBiomed Anal 15, 469 (1997).

Although the present invention has been described in considerable detailwith reference to certain preferred embodiments thereof, other versionsare possible. Therefore, the spirit and scope of the appended claimsshould not be limited to the preferred embodiments contained herein.

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1. A method for the treatment of a beta-Amyloid protein-induced oculardisease comprising administering to a subject suffering from abeta-Amyloid protein induced ocular disease a therapeutically effectiveamount of a composition comprising a member selected from a) a naturalor synthetic turmeric compound having anti-βA peptide activity; b) anatural or synthetic ginkgo biloba compound having anti-βA peptideactivity; and c) a natural or synthetic ginger compound having anti-βApeptide activity; d) a natural or synthetic sage compound having anti-βApeptide activity; and e) a natural or synthetic rosemary compound havinganti-βA peptide activity.
 2. The method of claim 1 for the treatment ofa beta-Amyloid protein-induced ocular disease comprising administeringto a subject suffering from the beta-Amyloid protein-induced oculardisease a therapeutically effective amount of a compound selected fromthe group consisting of: a) a compound having the formula (I):

b) a compound having the formula (II):

c) a compound having the formula (III):

or pharmaceutically acceptable salts or esters thereof, wherein: thedotted configuration

is optionally a single bond or a double bond (cis or trans) or a triplebond; Z is a representation of isosteric variation in which Z isselected from O, S, NH, NR₆₀, where R₆₀ is alkyl, alkenyl, or alkynyl;R₁ is selected from the group consisting of H, OH, OMe, CX_(m), F, Cl,Br, I, and OR₅₀ wherein R₅₀ is alkyl, alkenyl, or alkynyl, and X is F,Cl, Br, I, and m=1˜3; R₂ is selected from the group consisting of H, OH,OMe, CX_(m), F, Cl, Br, I, and OR₅₀ wherein R₅₀ is alkyl, alkenyl, oralkynyl, and X is F, Cl, Br, I, and m=1˜3; R₃ is selected from the groupconsisting of H, OH, OMe, CX_(m), F, Cl, Br, I, and OR₅₀ wherein R₅₀ isalkyl, alkenyl, or alkynyl, and X is F, Cl, Br, I, and m=1˜3; R₄ isselected from the group consisting of H, OH, OMe, CX_(m), F, Cl, Br, I,and OR₅₀ wherein R₅₀ is alkyl, alkenyl, or alkynyl, and X is F, Cl, Br,I, and m=1˜3; R₅ is selected from the group consisting of H, OH, OMe,CX_(m), F, Cl, Br, I, and OR₅₀ wherein R₅₀ is alkyl, alkenyl, oralkynyl, and X is F, Cl, Br, I, and m=1˜3; R₆ is selected from the groupconsisting of H, OH, OMe, CX_(m), F, Cl, Br, I, and OR₅₀ wherein R₅₀ isalkyl, alkenyl, or alkynyl, and X is F, Cl, Br, I, and m=1˜3; R₇ isselected from the group consisting of H, OH, OMe, CX_(m), F, Cl, Br, I,and OR₅₀ wherein R₅₀ is alkyl, alkenyl, or alkynyl, and X is F. Cl, Br,I, and m=1˜3; R₈ is selected from the group consisting of H, OH, OMe,CX_(m), F, Cl, Br, I, and OR₅₀ wherein R₅₀ is alkyl, alkenyl, oralkynyl, and X is F, Cl, Br, I, and m=1˜3; R₉ is selected from the groupconsisting of H, OH, OMe, CX_(m), F, Cl, Br, I, and OR₅₀ wherein R₅₀ isalkyl, alkenyl, or alkynyl, and X is F, Cl, Br, I, and m=1˜3.
 3. Themethod according to claim 1 wherein the disease is characterized bybeta-Amyloid protein induced cytotoxicity against retinal cells.
 4. Themethod according to claim 1 wherein the beta-Amyloid protein induceddisease is selected from the group consisting of age-related maculardegeneration (AMD) and glaucoma.
 5. The method of claim 1 for thetreatment of a beta-Amyloid protein-induced ocular disease comprisingadministering to a subject suffering from the beta-Amyloidprotein-induced ocular disease a therapeutically effective amount of acompound having the formula (IV):

or a pharmaceutically acceptable salt or ester thereof, wherein: R isselected from the group consisting of higher alkyl, higher alkenyl, andhigher alkynyl.
 6. The method according to claim 6 wherein: R is

 and n is 1-7, and dotted line is optionally a single bond, a doublebond (cis or trans), or a triple bond, or having more than one double ortriple bond consisting of; for example;

And R is also selected from the group consisting of alkyl, alkenyl, andalkynyl; for example;

and y is 1-9, or having more than one double bond (cis or trans), ortriple bond consisting of; for example;

wherein the dotted line configuration is optionally a single bond (cisor trans), or a triple bond, wherein the alkyl, alkenyl, and alkynylgroup is selected from ethers and/or thioethers or amines; for example;

wherein z=O, S, NR_(n), where R=alkyl, alkenyl, alynyl groups; and n=1or
 2. 7. The method of claim 1 for the treatment of a beta-Amyloidprotein-induced ocular disease comprising administering to a subjectsuffering from the beta-Amyloid protein-induced ocular disease atherapeutically effective amount of a compound having the formula (V):

or a pharmaceutically acceptable salt or ester thereof, wherein: thedotted configuration

is optionally a single bond, a double bond (cis or trans), or a triplebond; R₁₀ is selected from the group consisting of H, OH, OMe, CX_(m),F, Cl, Br, I, and OR₅₀ wherein R₅₀ is alkyl, alkenyl, or alkynyl, and Xis F, Cl, Br, I, and m=1˜3; R₁₁ is selected from the group consisting ofH, OH, OMe, CX_(m), F, Cl, Br, I, and OR₅₀ wherein R₅₀ is alkyl,alkenyl, or alkynyl, and X is F, Cl, Br, I, and m=1˜3;

and y is 1-9, or having more than one double bond (cis or trans), ortriple bond consisting of; for example;

wherein the dotted line configuration is optionally a single bond (cisor trans), or a triple bond, wherein the alkyl, alkenyl, and alkynylgroup is selected from ethers and/or thioethers or amines; for example;

wherein z=O, S, NR_(n) where R=alkyl, alkenyl, alynyl groups; and n=1 or2.
 8. The method of claim 1 for the treatment of a beta-Amyloidprotein-induced ocular disease comprising administering to a subjectsuffering from the beta-Amyloid protein-induced ocular disease atherapeutically effective amount of a compound having a formula (VI):

or a pharmaceutically acceptable salt or ester thereof, wherein: thedotted configuration

is optionally a single bond or a double bond or a triple bond; R₁₃ isselected from the group consisting of OH, OMe, OR′, and X wherein R′ isalkyl, alkenyl, or alkynyl, and X is F, Cl, Br, or I; R₁₄ is selectedfrom the group consisting of H, OH, OMe, and OR′ wherein R′ is alkyl,alkenyl, or alkynyl; and R₁₅ is selected from the group consisting ofalkyl, alkenyl, and alkynyl; for example;

and y is 1-9, or having more than one double bond (cis or trans), ortriple bond consisting of; for example;

wherein the dotted line configuration is optionally a single bond (cisor trans), or a triple bond, wherein the alkyl, alkenyl, and alkynylgroup is selected from ethers and/or thioethers or amines; for example;

wherein z=O, S, NR_(n) where R=alkyl, alkenyl, alynyl groups; and n=1 or2.
 9. The method of claim 1 for the treatment of a beta-amyloidprotein-induced ocular disease comprising administering to a subjectsuffering from the beta-amyloid protein-induced ocular disease atherapeutically effective amount of a compound selected from the groupconsisting of: a) a compound having the formula (VII):

b) a compound having the formula (VIII):

c) a compound having the formula (IX):


10. The method of claim 1 wherein the composition comprises compoundshaving anti-βA peptide activity from more than one of groups a) throughe).
 11. The method of claim 1 wherein the composition further comprisesone or more ingredients selected from the group consisting ofcholinergic agents, oral carbonic anhydrase inhibitors, alpha-2adrenergic agonists, prostaglandin agonists, carotenoids, lutein andzeaxanthin.